Substituted bicylic thiophene derivatives, compositions containing such compounds and methods of use

ABSTRACT

The present invention addresses substituted thiophene derivatives, as well as compositions containing such compounds and methods of treatment. The compounds in the present invention are glucagon antagonists. The compounds block the action of glucagon at its receptor and thereby decrease the levels of plasma glucose providing a treatment of diabetes.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is the National Stage of International Application No.PCT/US03/28033, filed 8 Sep. 2003, which claims the benefit under 35U.S.C. 119(e) of U.S. Provisional Application No. 60/410,145, filed 12Sept. 2002.

BACKGROUND OF THE INVENTION

The present invention relates to substituted thiophene derivatives,compositions containing such compounds and methods of treating type 2diabetes mellitus.

Diabetes refers to a disease process derived from multiple causativefactors and is characterized by elevated levels of plasma glucose(hyperglycemia) in the fasting state or following glucose administrationduring an oral glucose tolerance test. Frank diabetes mellitus (e.g., ablood glucose level ≧126 mg/dL in a fasting state) is associated withincreased and premature cardiovascular morbidity and mortality, and isrelated directly and indirectly to various metabolic conditions,including alterations of lipid, lipoprotein and apolipoproteinmetabolism.

Patients with non-insulin dependent diabetes mellitus (type 2 diabetesmellitus), approximately 95% of patients with diabetes mellitus,frequently display elevated levels of serum lipids, such as cholesteroland triglycerides, and have poor blood-lipid profiles, with high levelsof LDL-cholesterol and low levels of HDL-cholesterol. Those sufferingfrom Type 2 diabetes mellitus are thus at an increased risk ofdeveloping macrovascular and microvascular complications, includingcoronary heart disease, stroke, peripheral vascular disease,hypertension (for example, blood pressure ≧130/80 mmHg in a restingstate), nephropathy, neuropathy and retinopathy.

Patients having type 2 diabetes mellitus characteristically exhibitelevated plasma insulin levels compared with nondiabetic patients; thesepatients have developed a resistance to insulin stimulation of glucoseand lipid metabolism in the main insulin-sensitive tissues (muscle,liver and adipose tissues). Thus, Type 2 diabetes, at least early in thenatural progression of the disease is characterized primarily by insulinresistance rather than by a decrease in insulin production, resulting ininsufficient uptake, oxidation and storage of glucose in muscle,inadequate repression of lipolysis in adipose tissue, and excessglucose-production and secretion by the liver. The net effect ofdecreased sensitivity to insulin is high levels of insulin circulatingin the blood without appropriate reduction in plasma glucose(hyperglycemia). Hyperinsulinemia is a risk factor for developinghypertension and may also contribute to vascular disease.

Glucagon serves as the major regulatory hormone attenuating the effectof insulin in its inhibition of liver gluconeogenesis and is normallysecreted by α-cells in pancreatic islets in response to falling bloodglucose levels. The hormone binds to specific receptors in liver cellsthat triggers glycogenolysis and an increase in gluconeogenesis throughcAM-mediated events. These responses generate glucose (e.g. hepaticglucose production) to help maintain euglycemia by preventing bloodglucose levels from falling significantly.

In addition to elevated levels of circulating insulin, type II diabeticshave elevated levels of plasma glucagon and increased rates of hepaticglucose production. Antagonists of glucagon are useful in improvinginsulin responsiveness in the liver, decreasing the rate ofgluconeogenesis and lowering the rate of hepatic glucose outputresulting in a decrease in the levels of plasma glucose.

SUMMARY OF INVENTION

The present invention is directed to a compound represented by formulaI:

or a pharmaceutically acceptable salt or solvate thereof wherein:

X is NR⁴ or CR⁵R⁶;

R¹ is selected from the group consisting of: H, C₁₋₁₀alkyl,C₃₋₇cycloalkyl and Aryl, said alkyl, cycloalkyl and Aryl beingoptionally substituted with 1-4 substituents independently selected fromR¹³;

R² is selected from the group consisting of: R¹ as defined above,—C(O)₂R⁷ and —CONR⁷R⁸;

m and n are selected from 0, 1, 2 and 3, such that the sum of m and n is2 or 3, and when m is greater than 1, no more than one R¹ and no morethan one R² can be other than H;

R³ is selected from the group consisting of: C₁₋₁₀alkyl, C₃₋₇cycloalkyland Aryl, said alkyl, cycloalkyl and Aryl being optionally substitutedwith 14 substituents selected from R¹³, such that when R³ representsC₁₋₁₀ alkyl substituted with one R¹³ group, and R¹³ represents halo, R¹,R², R⁵ and R⁶ do not represent C₁₋₃alkyl;

R⁴ is selected from the group consisting of: C₃₋₁₀ alkyl, C₃₋₇cycloalkyl, Aryl, HAR, Hetcy, C(O)C₅₋₁₀ alkyl, C(O)C₃₋₇ cycloalkyl,C(O)-Aryl, C(O)—HAR, C(O)-Hetcy, CONR⁹R¹⁰, CO₂R⁹ and SO₂R⁹, the alkyl,cycloalkyl, Aryl, HAR and Hetcy groups and portions being optionallysubstituted with 1-4 substituents selected from R¹³;

one of R⁵ and R⁶ is selected from the group consisting of NR¹¹R¹²,NR¹¹CORR¹², NR¹¹CO₂R¹² and NR¹¹S(O)₂R¹², and the other represents R¹,HAR, Hetcy or OR¹¹, said HAR and Hetcy being optionally substituted with1-4 substituents selected from R¹³,

R⁷, R¹⁰ and R¹¹ are selected from the group consisting of: R¹ as definedabove, HAR and Hetcy, said HAR and Hetcy being optionally substitutedwith 1-4 substituents selected from R¹³;

R⁸, R⁹ and R¹² are selected from the group consisting of: C₁₋₁₀alkyl,C₃₋₇cycloalkyl, Aryl, HAR and Hetcy, said alkyl, cycloalkyl, Aryl, HARand Hetcy being optionally substituted with 1-4 substituents selectedfrom R¹³;

-   -   or alternatively, R⁷, R⁸, R⁹ and R¹⁰ are as defined above, and        R¹¹ and R¹² are taken together with the atoms to which they are        attached along with any intervening atoms and represent a 5-8        membered ring optionally containing 1-2 heteroatoms selected        from O, S and N, and optionally substituted with 14 substituents        selected from R¹³;

each R¹³ is selected from the group consisting of: halo, NR¹⁴R¹⁵,C₁₋₄alkyl, C₃₋₇ cycloalkyl, Aryl, HAR, Hetcy, CF₃, OCF₃, OR¹⁵, NO₂,S(O)_(x)R¹⁴, SR¹⁴, S(O)_(x)NR¹⁴R¹⁵, O(CR¹⁶R¹⁷)NR¹⁴R¹⁵, C(O)R¹⁴, CO₂R¹⁵,CO₂(CR¹⁶R¹⁷)_(y)CONR¹⁴R¹⁵, OC(O)R¹⁴, CN, C(O)NR¹⁴R¹⁵, NR¹⁵C(O)R¹⁴,NR¹⁵C(O)OR¹⁴, NR¹⁵C(O)NR¹⁶R¹⁴ and CR¹⁵(N—OR¹⁴), wherein x is 1 or 2, andy is an integer from 1-4,

said alkyl, cycloalkyl, Aryl, HAR and Hetcy being optionally substitutedwith 1-4 substituents selected from R¹⁸;

R¹⁴, R¹⁵, R¹⁶ and R¹⁷ are independently selected from the groupconsisting of: H, C₁₋₁₀alkyl, C₃₋₇cycloalkyl, Aryl and Ar—C₁₋₁₀alkyl;

and each R¹⁸ is independently selected from the group consisting of:halogen, CN, C₁₋₄alkyl, OH, CF₃, Aryl, Aryloxy, CO₂H and CO₂C₁₋₄ alkyl,said Aryl and the Aryl portion of Aryloxy being optionally substitutedwith up to 4 halo groups, and up to 2 C₁₋₄ alkyl, OH, CF₃ or CN groups.

DETAILED DESCRIPTION OF THE INVENTION

The invention is described herein in detail using the terms definedbelow unless otherwise specified.

“Alkyl”, as well as other groups having the prefix “alk”, such asalkoxy, alkanoyl and the like, means carbon chains which may be linear,branched, or cyclic, or combinations thereof, containing the indicatednumber of carbon atoms. If no number is specified, 1-10 carbon atoms areintended for linear or branched alkyl groups. Examples of alkyl groupsinclude methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl,pentyl, hexyl, heptyl, octyl, nonyl and the like. Cycloalkyl is a subsetof alkyl; if no number of atoms is specified, 3-10 carbon atoms areintended, forming 1-3 carbocyclic rings that are fused. “Cycloalkyl”also includes monocyclic rings fused to an aryl group in which the pointof attachment is on the non-aromatic portion. Examples of cycloalkylinclude cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,tetrahydronaphthyl, decahydronaphthyl, indanyl and the like.

“Alkenyl” means carbon chains which contain at least one carbon-carbondouble bond, and which may be linear or branched or combinationsthereof. Examples of alkenyl include vinyl, allyl, isopropenyl,pentenyl, hexenyl, heptenyl, 1-propenyl, 2-butenyl, 2-methyl-2-butenyl,and the like.

“Alkynyl” means carbon chains which contain at least one carbon-carbontriple bond, and which may be linear or branched or combinationsthereof. Examples of alkynyl include ethynyl, propargyl,3-methyl-1-pentynyl, 2-heptynyl and the like.

“Aryl” (Ar) means mono- and bicyclic aromatic rings containing onlycarbon atoms. Examples of aryl include phenyl and naphthyl.

“Heteroaryl” (HAR) means a mono- or bicyclic aromatic ring or ringsystem containing at least one heteroatom selected from O, S and N, witheach ring containing 5 to 6 atoms. Examples include pyrrolyl,isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, oxadiazolyl,thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl,triazinyl, thienyl, pyrimidyl, pyridazinyl, pyrazinyl, benzoxazolyl,benzothiazolyl, benzimidazolyl, benzofuranyl, benzothiophenyl,furo(2,3-b)pyridyl, quinolyl, indolyl, isoquinolyl and the like.Heteroaryl also includes aromatic heterocyclic groups fused toheterocycles that are non-aromatic or partially aromatic, and aromaticheterocyclic groups fused to cycloalkyl rings.

“Heterocyclyl” (Hetcy) means mono- and bicyclic saturated rings and ringsystems containing at least one heteroatom selected from N, S and O,each of said ring having from 3 to 10 atoms in which the point ofattachment may be carbon or nitrogen. Examples of “heterocyclyl” includepyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl,2,3-dihydrofuro(2,3-b)pyridyl, benzoxazinyl, tetrahydrohydroquinolinyl,tetrahydroisoquinolinyl, dihydroindolyl, and the like. The term alsoincludes partially unsaturated monocyclic rings that are not aromatic,such as 2- or 4-pyridones attached through the nitrogen orN-substituted-(1H,3H)-pyrimidine-2,4-diones (N-substituted uracils).

“Halogen” (Halo) includes fluorine, chlorine, bromine and iodine.

The present invention is directed to a compound represented by formulaI:

or a pharmaceutically acceptable salt or solvate thereof wherein:

X is NR⁴ or CR⁵R⁶;

R¹ is selected from the group consisting of: H, C₁₋₁₀alkyl,C₃₋₇cycloalkyl and Aryl, said alkyl, cycloalkyl and Aryl beingoptionally substituted with 1-4 substituents independently selected fromR¹³;

R² is selected from the group consisting of: R¹ as defined above,—C(O)₂R⁷ and —CONR⁷R⁸;

m and n are selected from 0, 1, 2 and 3, such that the sum of m and n is2 or 3, and when m is greater than 1, no more than one R¹ and no morethan one R² can be other than H;

R³ is selected from the group consisting of: C₁₋₁₀alkyl, C₃₋₇cycloalkyland Aryl, said alkyl, cycloalkyl and Aryl being optionally substitutedwith 1-4 substituents selected from R¹³, such that when R³ representsC₁₋₁₀alkyl substituted with one R¹³ group, and R¹³ represents halo, R¹,R², R⁵ and R⁶ do not represent C₁₋₃alkyl;

R⁴ is selected from the group consisting of: C₃₋₁₀ alky, C₃₋₇cycloalkyl, Aryl, HAR, HetCy, C(O)C₅₋₁₀ alkyl, C(O)C₃₋₇ cycloalkyl,C(O)-Aryl, C(O)—HAR, C(O)-Hetcy, CONR⁹R¹⁰, CO₂R⁹ and SO₂R⁹, the alkyl,cycloalkyl, Aryl, HAR and Hetcy groups and portions being optionallysubstituted with 14 substituents selected from R¹³;

one of R⁵ and R⁶ is selected from the group consisting of NR¹¹R¹²,NR¹¹COR¹², N¹¹CO₂R¹² and NR¹¹S(O)₂R¹², and the other represents R¹, HAR,Hetcy or OR¹¹, said HAR and Hetcy being optionally substituted with 14substituents selected from R¹³,

R⁷, R¹⁰ and R¹¹ are selected from the group consisting of: R¹ as definedabove, HAR and Hetcy, said HAR and Hetcy being optionally substitutedwith 14 substituents selected from R¹³;

R⁸, R⁹ and R¹² are selected from the group consisting of: C₁₋₁₀alkyl,C₃₋₇cycloalkyl, Aryl, HAR and Hetcy, said alkyl, cycloalkyl, Aryl, HARand Hetcy being optionally substituted with 1-4 substituents selectedfrom R¹³;

or alternatively, R⁷, R⁸, R⁹ and R¹⁰ are as defined above, and R¹¹ andR¹² are taken together with the atoms to which they are attached alongwith any intervening atoms and represent a 5-8 membered ring optionallycontaining 1-2 heteroatoms selected from O, S and N, and optionallysubstituted with 14 substituents selected from R¹³;

each R¹³ is selected from the group consisting of: halo, NR¹⁴R¹⁵,C₁₄alkyl, C₃₋₇ cycloalkyl, Aryl, HAR, Hetcy, CF₃, OCF₃, OR¹⁵, NO₂,S(O)_(x)R¹⁴, SR¹⁴, S(O)_(x)NR¹⁴R¹⁵, O(CR¹⁶R¹⁷)_(y)NR¹⁴R¹⁵, C(O)R¹⁴,CO₂R¹⁵, CO₂(CR¹⁶R¹⁷)_(y)CONR¹⁴R¹⁵) OC(O)R¹⁴, CN, C(O)NR¹⁴R¹⁵,NR¹⁵C(O)R¹⁴, NR¹⁵C(O)OR¹⁴, NR¹⁵C(O)NR¹⁶R¹⁴ and CR¹⁵(N—OR¹⁴), wherein xis 1 or 2, and y is an integer from 14,

said alkyl, cycloallcyl, Aryl, HAR and Hetcy being optionallysubstituted with 1-4 substituents selected from R¹⁸;

R¹⁴, R¹⁵, R¹⁶ and R¹⁷ are independently selected from the groupconsisting of: H, C₁₋₁₀alkyl, C₃₋₇cycloalkyl, Aryl and Ar—C₁₋₁₀alkyl;

and each R¹⁸ is independently selected from the group consisting of:halogen, CN, C₁₋₄alkyl, OH, CF₃, Aryl, Aryloxy, CO₂H and CO₂C₁₋₄ alkyl,said Aryl and the Aryl portion of Aryloxy being optionally substitutedwith up to 4 halo groups, and up to 2 C₁₋₄ alkyl, OH, CF₃ or CN groups.

In an aspect of the invention that is of particular interest, a compoundof formula I is disclosed wherein R¹ is selected from the groupconsisting of: H, C₁₋₁₀alkyl, C₃₋₄ cycloalkyl and phenyl, said alkyl andphenyl being optionally substituted with 1-3 substituents selected fromR¹³. Within this aspect of the invention, all other variables are asoriginally defined.

In another aspect of the invention that is of particular interest, acompound of formula I is disclosed wherein R² is H. Within this aspectof the invention, all other variables are as originally defined.

In another aspect of the invention that is of particular interest, acompound of formula I is disclosed wherein m is 0 and n is 2 or 3, or mis 1 and n is 1 or 2. Thus, the sum of m and n is 2 or 3. Within thisaspect of the invention, all other variables are as originally defined.

In another aspect of the invention that is of particular interest, acompound of formula I is disclosed wherein R³ is C₃₋₁₀ alkyl optionallysubstituted with 14 substituents selected from R such that when R³ issubstituted with one R¹³ group, and R¹³ represents halo, R¹, R², R⁵ andR⁶ do not represent C₁₋₃alkyl. Within this aspect of the invention, allother variables are as originally defined.

More particularly, an aspect of the invention that is of particularinterest relates to a compound of formula I wherein R³ represents C₃₋₅alkyl, optionally substituted with 1-4 R¹³ groups. Within this aspect ofthe invention, all other variables are as originally defined.

In another aspect of the invention that is of particular interest, acompound of formula I is disclosed wherein R⁴ is selected from the groupconsisting of: C₅₋₁₀ alkyl, C₃₋₆cycloalkyl, phenyl, HAR, Hetcy,C(O)C₅₋₁₀alkyl, C(O)C₃₋₆cycloalkyl and CO₂R⁹, the alkyl, cycloalkyl and,Aryl groups and portions, phenyl, HAR and Hetcy being optionallysubstituted with 1-4 substituents selected from R¹³, and R⁹ representingC₁₋₁₀alkyl, C₃₋₇cycloalkyl, Aryl, HAR or Hetcy, said alkyl, cycloalkyl,Aryl groups and portions, HAR and Hetcy being optionally substitutedwith 1-4 R¹³ groups. Within this aspect of the invention, all othervariables are as originally defined.

In another aspect of the invention that is of particular interest, acompound of formula I is disclosed wherein X represents CR⁵R⁶, R⁵ isNR¹¹R¹², and R⁶ is selected from the group consisting of: R¹, HAR, Hetcyand OR¹¹, wherein R¹ is as originally defined, R¹¹ is R¹ or HAR, and R¹²is C₁₋₆alkyl, Aryl or HAR, said Aryl and HAR being optionallysubstituted with 1-4 R¹³ groups,

or R¹¹ and R¹² are taken in combination with the atom to which they areattached and represent a 5-6 membered ring optionally substituted with1-2 R¹³ groups. Within this aspect of the invention, all other variablesare as originally defined.

In another aspect of the invention, a compound of formula I is disclosedwherein R¹³ is selected from the group consisting of: halo, C₁₋₄alkyl,C₃₋₇cycloalkyl, Aryl, HAR, Hetcy, and OR¹⁵ wherein R¹⁵ is H,

said alkyl, cycloalkyl, Aryl, HAR and Hetcy being optionally substitutedwith 14 substituents selected from R¹⁸ and

R¹⁸ is halo, C₁₋₄alkyl, Aryl or CO₂C₁₋₄ alkyl.

Within this aspect of the invention, all other variables are asoriginally defined.

In yet another aspect of the invention that is of particular interest, acompound of formula I is disclosed wherein:

R¹ is selected from the group consisting of: H, C₁₋₁₀alkyl, C₃₋₆cycloalkyl and phenyl, said alkyl and phenyl being optionallysubstituted with 1-3 substituents selected from R¹³;

R² is H;

m is 0 and n is 2 or 3, or m is 1 and n is 1 or 2, such that the sum ofm and n is 2 or 3;

R³ is C₃₋₁₀ alkyl optionally substituted with 14 substituents selectedfrom R¹³, such that when R³ is substituted with one R¹³ group, and R¹³represents halo, R¹, R², R⁵ and R⁶ do not represent C₁₋₃alkyl;

R⁴ is selected from the group consisting of: C₅₋₁₀ alkyl, C₃₋₆cycloalkyl, phenyl, HAR, Hetcy, C(O)C₅₋₁₀alkyl, C(O)C₃₋₆ cycloalkyl andCO₂R⁹, the alkyl, cycloalkyl and, Aryl groups and portions, phenyl, HARand Hetcy being optionally substituted with 1-4 substituents selectedfrom R¹³, and R⁹ representing C₁₋₁₀alkyl, C₃₋₇cycloalkyl, Aryl, HAR orHetcy, said alkyl, cycloalkyl, Aryl groups and portions, HAR and Hetcybeing optionally substituted with 14 R¹³ groups;

X represents CR⁵R⁶, R⁵ is NR¹¹R¹², and R⁶ is selected from the groupconsisting of: R¹, HAR, Hetcy and OR¹¹, wherein R¹ is as originallydefined, R¹¹ is R¹ or HAR, and R¹² is C₁₋₆alkyl, Aryl or HAR, said Aryland HAR being optionally substituted with 1-4 R¹³ groups,

or R¹¹ and R¹² are taken in combination with the atom to which they areattached and represent a 5-6 membered ring optionally substituted with1-2 R¹³ groups;

R¹³ is selected from the group consisting of: halo, C₁₋₄alkyl,C₃₋₇cycloalkyl, Aryl, HAR, Hetcy, and OR¹⁵ wherein R¹⁵ is H,

said alkyl, cycloalkyl, Aryl, HAR and Hetcy being optionally substitutedwith 1-4 substituents selected from R¹⁸ and

R¹⁸ is halo, C₁₋₄alkyl, Aryl or CO₂C₁₋₄ alkyl.

Species falling within the scope of the present invention that are ofparticular interest include the following:

-   tert-butyl    3-cyano-2-[(2-ethylbutanoyl)amino]-5,6-dihydrothieno[2,3-b]pyridine-7(4H)-carboxylate;-   N-(3-cyano-7-isobutyl4,5,6,7-tetrahydrothieno[2,3-b]pyridin-2-yl)-2-ethylbutanamide;-   N-(3-cyano-7-isopropyl-4,5,6,7-tetrahydrothieno[2,3-b]pyridin-2-yl)-2-ethylbutanamide;-   N-{6-[(4′-chloro-1,1′-biphenyl-4-yl)methyl]-3-cyano-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl}-2-ethlylbutanamide;-   N-[3-cyano-6-(4-phenoxybenzyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-2-ethylbutanamide;-   N-{6-[4-(4-chlorophenoxy)benzyl]-3-cyano-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl}-2-ethylbutanamide;-   N-[3-cyano-6-(3-phenoxybenzyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-2-ethylbutanamide;-   N-(3-cyano-6-{[1-(2,4-dichlorophenyl)cyclopropyl]carbonyl}4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-2-ethylbutanamide;-   N-{3-cyano-6-[(2,4-dichlorobenzyl)amino]4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(cyclopropylmethyl)(2,4-dichlorobenzyl)amino]4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(2,4-dichlorobenzyl)(isopropyl)amino]4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(2,4-dichlorobenzyl)(isopentyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(2,4-dichlorobenzyl)(3,3-dimethylbutyl)amino]4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(2,4-dichlorobenzyl)(isobutyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(2,4-dichlorobenzyl)(2-ethylbutyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-(3-cyano-6-{(2,4-dichlorobenzyl)[(4,5-dimethyl-2-furyl)methyl]amino}-4,5,6,7-tetrahydro-1-benzothien-2-yl)-2-ethylbutanamide;-   N-{3-cyano-6-[(2,4-dichlorobenzyl)(3-phenylpropyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{6-[(1-benzofuran-2-ylmethyl)(2,4-dichlorobenzyl)amino]-3-cyano-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(2,4-dichlorobenzyl)(3,3,3-trifluoropropyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(2,4-dichlorobenzyl)(4-fluorobenzyl)amino)-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(2,4-dichlorobenzyl)(tetrahydrofuran-2-ylmethyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-(3-cyano-6-{(2,4-dichlorobenzyl)[(5-methyl-2-furyl)methyl]amino}-4,5,6,7-tetrahydro-1-benzothien-2-yl)-2-ethylbutanamide;-   tert-butyl    (2S)-2-{[{3-cyano-2-[(2-ethylbutanoyl)amino]-4,5,6,7-tetrahydro-1-benzothien-6-yl}(2,4-dichlorobenzyl)amino]methyl}pyrrolidine-1-carboxylate;-   N-{3-cyano-6-[(3,4-dichlorobenzyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(3,4-dichlorobenzyl)(methyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-(3-cyano-6-{[(2-phenyl-1,3-thiazol-5-yl)methyl]amino}-4,5,6,7-tetrahydro-1-benzothien-2-yl)-2-ethylbutanamide;-   N-(3-cyano-6-{methyl[(2-phenyl-1,3-thiazol-5-yl)methyl]amino}-4,5,6,7-tetrahydro-1-benzothien-2-yl)-2-ethylbutanamide;-   N-(3-cyano-6-{[(2-phenyl-1,3-thiazol-4-yl)methyl]amino-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-(3-cyano-6-{methyl[(2-phenyl-1,3-thiazol-4-yl)methyl]amino}4,5,6,7-tetrahydro-1-benzothien-2-yl)-2-ethylbutanamide;-   N-{3-cyano-6-(1,2,3,4-tetrahydronaphthalen-1-ylamino)-4,5,6,7-tetrahydro-1-benzothien-2-yl]-2-ethylbutanamide;-   N-{3-cyano-6-[methyl(1,2,3,4-tetrahydronaphthalen-1-yl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(2,3-dihydro-1H-inden-1-ylmethyl)amino]}4,5,6,7-tetrahydrol-benzothien-2-yl}-2-ethylbutanamide;-   N-{3-cyano-6-[(2,3-dihydro-1H-inden-1-ylmethyl)(methyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-{6-[(2-chlorobenzyl)amino]-3-cyano-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide-   N-{6-[(2-chlorobenzyl)(methyl)amino]-3-cyano-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-(6-{[1-(4-bromophenyl)ethyl]amino}-3-cyano-4,5,6,7-tetrahydro-1-benzothien-2-yl)-2-ethylbutanamide;-   N-{6-[[1-(4-bromophenyl)ethyl](methyl)amino]-3-cyano-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide;-   N-[3-cyano-6-(3-phenylpyrrolidin-1-yl)-4,5,6,7-tetrahydro-1-benzothien-2-yl]-2-ethylbutanamide;-   N-[3-cyano-6-(4-phenylpiperazin-1-yl)-4,5,6,7-tetrahydro-1-benzothien-2-yl]-2-ethylbutanamide;-   N-{3-cyano-2-[(2-ethylbutanoyl)amino]-4,5,6,7-tetrahydro-1-benzothien-6-yl}-N-(2,4-dichlorobenzyl)-3,3-dimethylbutanamide;-   N-{3-cyano-2-[(2-ethylbutanoyl)amino]-4,5,6,7-tetrahydro-1-benzothien-6-yl}-N-[1-(hydroxymethyl)-2,2-dimethylpropyl)cyclopropanecarboxamide;-   N-{3-cyano-2-[(2-ethylbutanoyl)amino]-4,5,6,7-tetrahydro-1-benzothien-6-yl}-N-[1-(hydroxymethyl)-2,2-dimethylpropyl]-3,3-dimethylbutanamide;-   N-{3-cyano-2-[(2-ethylbutanoyl)amino]-4,5,6,7-tetrahydro-1-benzothien-6-yl}-N-[1-(hydroxymethyl)-2,2-dimethylpropyl]cyclopentanecarboxamide;-   N-{3-cyano-2-[(2-ethylbutanoyl)amino]-4,5,6,7-tetrahydro-1-benzothien-6-yl}-N-[1-(hydroxymethyl)-2,2-dimethylpropyl]benzamide    and-   N-{3-cyano-2-[(2-ethylbutanoyl)amino]4,5,6,7-tetrahydro-1-benzothien-6-yl}-N-[1-(hydroxymethyl)-2,2-dimethylpropyl]cyclohexanecarboxamide.

The invention further includes a pharmaceutical composition which iscomprised of a compound of formula I in combination with apharmaceutically acceptable carrier.

Also included is a method of treating type 2 diabetes mellitus in amammalian patient in need of such treatement, comprising administeringto said patient a compound of formula I in an amount that is effectiveto treat type 2 diabetes mellitus.

Also included is a method of preventing or delaying the onset of type 2diabetes mellitus in a mammalian patient in need thereof, comprisingadministering to said patient a compound of formula I in an amount thatis effective to prevent or delay the onset of type 2 diabetes mellitus.

Also included in a method of treating, preventing or delaying the onsetof diseases or conditions that are associated with type 2 diabetesmellitus. Examples include diseases and conditions selected from thegroup consisting of: dyslipidemias, such as elevated levels ofcholesterol, triglycerides or low density lipoproteins (LDL), low levelsof high density lipoprotein (HDL), microvascular or macrovascularchanges and the sequellae of such conditions, such as coronary heartdisease, stroke, peripheral vascular disease, hypertension, renalhypertension, nephropathy, neuropathy and retinopathy. The methodentails administering to a type 2 diabetic patient, e.g., a humanpatient, an amount of a compound of formula I that is effective fortreating, preventing or delaying the onset of such diseases orconditions.

Optical Isomers—Diastereomers—Geometric Isomers—Tautomers

Many of the compounds of formula I contain one or more asymmetriccenters and thus occur as racemates and racemic mixtures, singleenantiomers, diastereomeric mixtures and individual diastereomers. Thepresent invention includes all such isomeric forms of the compounds, inpure form as well as in mixtures.

Some of the compounds described herein contain olefinic double bonds,and unless specified otherwise, are meant to include both E and Zgeometric isomers.

Some of the compounds described herein may exist with different pointsof attachment of hydrogen, referred to as tautomers. Such an example maybe a ketone and its enol form known as keto-enol tautomers. Theindividual tautomers as well as mixture thereof are encompassed withcompounds of Formula I.

Salts and Solvates

The term “pharmaceutically acceptable salts” refers to salts preparedfrom pharmaceutically acceptable substantially non-toxic bases or acidsincluding inorganic or organic bases and inorganic or organic acids, aswell as salts that can be converted into pharmaceutically acceptablesalts. Salts derived from inorganic bases include aluminum, ammonium,calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts,manganous, potassium, sodium, zinc, and the like. Particularly preferredare the ammonium, calcium, magnesium, potassium, and sodium salts. Saltsderived from pharmaceutically acceptable organic non-toxic bases includesalts of primary, secondary, and tertiary amines, substituted aminesincluding naturally occurring substituted amines, cyclic amines, andbasic ion exchange resins, such as ethylmorpholine, N-ethylpiperidine,glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobrornine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like.

When the compound of the present invention is basic, salts may beprepared from pharmaceutically acceptable non-toxic acids, includinginorganic and organic acids. Such acids include acetic, benzenesulfonic,benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic,glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic,mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic,phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid, andthe like. Particularly preferred are citric, hydrobromic, hydrochloric,maleic, phosphoric, sulfuric, and tartaric acids.

Solvates as used herein refers to the compound of formula I or a saltthereof, in association with a solvent, such as water. Representativeexamples include hydrates, hemihydrates, trihydrates and the like.

References to the compounds of Formula I include the pharmaceuticallyacceptable salts and solvates.

This invention relates to method of antagonizing or inhibiting theproduction or activity of glucagon, thereby reducing the rate ofgluconeogenesis and glycogenolysis, and the concentration of glucose inplasma.

The compounds of formula I can be used in the manufacture of amedicament for the prophylactic or therapeutic treatment of diseasestates in mammals caused by elevated levels of glucose.

Dose Rantes

The prophylactic or therapeutic dose of a compound of Formula I will, ofcourse, vary with the nature of the condition to be treated, theparticular compound selected and its route of administration. It willalso vary according to the age, weight and response of the individualpatient. In general, the daily dose range lie within the range of fromabout 0.001 mg to about 100 mg per kg body weight, preferably about 0.01mg to about 50 mg per kg, and more preferably 0.1 to 10 mg per kg, insingle or divided doses. It may be necessary to use dosages outside ofthese limits in some cases.

When intravenous or or oral administration is employed, a representativedosage range is from about 0.001 mg to about 100 mg (preferably from0.01 mg to about 10 mg) of a compound of Formula I per kg of body weightper day, and more preferably, about 0.1 mg to about 10 mg of a compoundof Formula I per kg of body weight per day.

Pharmaceutical Compositions

As mentioned above, the pharmaceutical composition comprises a compoundof Formula I and a pharmaceutically acceptable carrier. The term“composition” encompasses a product comprising the active and inertingredient(s), (pharmaceutically acceptable excipients) that make up thecarrier, as well as any product which results, directly or indirectly,from the combination, complexation or aggregation of any two or more ofthe ingredients, or from dissociation of one or more of the ingredients,or from other types of reactions or interactions between ingredients.Preferably the composition is comprised of a compound of formula I in anamount that is effective to treat, prevent or delay the onset of type 2diabetes mellitus, in combination with the pharmaceutically acceptablecarrier.

Any suitable route of administration may be employed for providing amammal, especially a human with an effective dosage of a compound of thepresent invention. For example, oral, rectal, topical, parenteral,ocular, pulmonary, nasal, and the like may be employed. Examples ofdosage forms include tablets, troches, dispersions, suspensions,solutions, capsules, creams, ointments, aerosols and the like, with oraltablets being preferred.

In preparing oral compositions, any of the usual pharmaceutical mediamay be employed, such as, for example, water, glycols, oils, alcohols,flavoring agents, preservatives, coloring agents and the like in thecase of oral liquids, e.g., suspensions, elixirs and solutions; orcarriers such as starches, sugars, microcrystalline cellulose, diluents,granulating agents, lubricants, binders, disintegrating agents and thelike in the case of oral solids, e.g., powders, capsules and tablets,with the solid oral preparations being preferred. Because of their easeof administration, tablets and capsules represent the most advantageousoral dosage unit forms. If desired, tablets may be coated by standardaqueous or nonaqueous techniques.

In addition to the common dosage forms set out above, the compounds ofFormula I may also be administered by controlled release means and/ordelivery devices such as those described in U.S. Pat. Nos. 3,845,770;3,916,899; 3,536,809; 3,598,123; 3,630,200 and 4,008,719.

Pharmaceutical compositions of the present invention suitable for oraladministration may be presented as discrete units such as capsules,cachets or tablets each containing a predetermined amount of the activeingredient, as a powder or granules or as a solution or a suspension inan aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion or awater-in-oil liquid emulsion. Such compositions may be prepared by anyof the methods of pharmacy but all methods include the step of bringinginto association the active ingredient with the carrier whichconstitutes one or more necessary ingredients. In general, thecompositions are prepared by uniformly and intimately admixing theactive ingredient with liquid carriers or finely divided solid carriersor both, and then, if necessary, shaping the product into the desiredpresentation. For example, a tablet may be prepared by compression ormolding, optionally with one or more accessory ingredients. Compressedtablets may be prepared by compressing in a suitable machine, the activeingredient in a free-flowing form such as powder or granules, optionallymixed with a binder, lubricant, inert diluent, surface active ordispersing agent. Molded tablets may be made by molding in a suitablemachine, a mixture of the powdered compound moistened with an inertliquid diluent. Desirably, each tablet contains from about 1 mg to about1 g of the active ingredient and each cachet or capsule contains fromabout 1 to about 500 mg of the active ingredient.

The following are examples of pharmaceutical dosage forms for thecompounds of Formula I:

Injectable Suspension (I.M.) mg/mL Tablet mg/tablet Compound of FormulaI 10 Compound of Formula I 25 Methylcellulose 5.0 MicrocrystallineCellulose 415 Tween 80 0.5 Povidone 14.0 Benzyl alcohol 9.0Pregelatinized Starch 43.5 Benzalkonium chloride 1.0 Magnesium Stearate2.5 Water for injection to make 1.0 mL Total 500 mg Capsule mg/capsuleAerosol Per canister Compound of Formula I 25 Compound of Formula I 24mg Lactose Powder 573.5 Lecithin, NF Liq. Conc. 1.2 mg MagnesiumStearate 1.5 Trichlorofluoromethane, NF 4.025 g Total 600 mgDichlorodifluoromethane, NF 12.15 gCombination Therapy

Compounds of Formula I may be used in combination with other drugs thatare used in the treatment/prevention/delaying the onset of type 2diabetes mellitus, as well as the diseases and conditions associatedwith type 2 diabetes mellitus, for which compounds of Formula I areuseful. Other drugs may be administered, by a route and in an amountcommonly used therefor, contemporaneously or sequentially with acompound of Formula I. When a compound of Formula I is usedcontemporaneously with one or more other drugs, a pharmaceuticalcomposition containing such other drugs in addition to the compound ofFormula I is preferred Accordingly, the pharmaceutical compositions ofthe present invention include those that also contain one or more otheractive ingredients, in addition to a compound of Formula I Examples ofother active ingredients that may be combined with a compound of FormulaI either administered separately or in the same pharmaceuticalcompositions, include, but are not limited to: (a) bis-guanides (e.g.,buformin, metformin, phenformin), (b) PPAR agonists (e.g., troglitazone,pioglitazone, rosiglitazone), (c) insulin, (d) somatostatin, (e)α-glucosidase inhibitors (e.g., voglibose, miglitol, acarbose), and (f)insulin secretagogues (e.g., acetohexamide, carbutamide, chlorpropamide,glibornuride, gliclazide, glimerpiride, glipizide, gliquidine,glisoxepid, glyburide, glyhexamide, glypinamide, phenbutamide,tolazamide, tolbutamide, tolcyclamide, nateglinide, repaglinide).

The weight ratio of the compound of the Formula I to the second activeingredient may be varied within wide limits and depends upon theeffective dope of each ingredient Generally, an effective dose of eachwill be used. Thus, for example, when a compound of the Formula I iscombined with a PPAR agonist the weight ratio of the compound of theFormula I to the PPAR agonist will generally range from about 1000:1 toabout 1:1000, preferably about 200:1 to about 1:200. Combinations of acompound of the Formula I and other active ingredients will generallyalso be within the aforementioned range, but in each case, an effectivedose of each active ingredient should be used.

Throughout the instant application, the following abbreviations are usedwith the following meanings unless otherwise indicated:

Bu = butyl Bn = benzyl BOC, Boc = t-butyloxycarbonyl CBZ, Cbz =Benzyloxycarbonyl DCC = Dicyclohexylcarbodiimide DCM = dichloromethaneDIEA = diisopropylethylamine DMF = N,-dimethylformamide DMAP =4-Dimethylaminopyridine Et = ethyl EtOAc = ethyl acetate EtOH = ethanoleq. = equivalent(s) FAB-mass spectrum = Fast atom bombardment-massspectroscopy HOAc = acetic acid HPLC = High pressure liquidchromatography HOBT, HOBt = Hydroxybenztriazole LAH = Lithium aluminumhydride Me = methyl PBS = phosphate buffer saline Ph = phenyl TFA =Trifluoroacetic acid THF = Tetrahydrofuran TMS = Trimethylsilane

Compounds of the present invention may be prepared according to themethodology outlined in the following Schemes.

Cyclic ketones such as 1, where X is CR⁵R⁶ from formula I, arecommercially available, known in the literature or may be convenientlyprepared by a variety of methods familiar to those skilled in the art.

In Scheme 1, a cyclic ketone 1 is condensed with malonitrile 2 in thepresence of sulfur (S₈) and a dialkylamine (e.g., morpholine) in ethanolaccording to methods described in the literature (S. Mukherjee and A.De, J. Chem. Res. 5, 295 (1994); M. S. Mahas et al. J. Chem. Soc. 1969,1937; A. De et al. J. Het. Chem. 29, 1213 (1992)) to afford2-amino-3-cyano-thiophene 3. Acylation of 3 with an appropriateanhydride or acid chloride in the presence of a trialkylamine (e.g.,diisopropylethylamine) according to published procedues (U. Sensfuss etal. Heteroat. Chem. 9, 529 (1998) will afford the amide represented byformula I.

It is recognized that when the cyclic ketone 1 is not a symmetricallysubstituted ketone, the product 3 may be formed as a mixture ofpositional isomers. These isomers may be separated at any stage in thesynthetic sequence by preparative thin layer chromatography, flashchromatography on silica gel as described by W. C. Still et al., J. Org.Chem., 43, 2923 (1978), or HPLC. Compounds that are purified by HPLC maybe isolated as the corresponding salt.

In some instances it may be necessary to carry out the thiophenesynthesis in two steps, as illustrated in Scheme 2.

A dicyano-alkene 4 is first prepared by condensation of a ketone such as1 and malonitrile. This intermediate is e d with sulfur (S₈) and adiallylamine (e.g., morpholine) in ethanol according to methods dearibedin the literature (A. Rajca and A Tisler, Monatch. Chem. 121, 697(1990); B. Naumamn et al., Pharmazie 53, 4 (1996)) to afford2-amino-3-cyanothiophene 3. Acylation of 3 with an appropriate anhydrideor acid chloride in the presence of a trialkylamine (e.g.,diisopropylethylamine) according to published procedures (U. Sensfuss etal. Heteroat. Chem. 9, 529 (1998) afford the thiopheneamide representedby formula I.

Intermediates such as 5 in Scheme 3, where n is an integer greater than0 and X is nitrogen are commercially available, known in the literatureor may be prepared by a variety of known methods.

In the schemes, R^(a), R^(b) and R^(c) are appended to reagents and canform part of a variable within the genus. Thus, for example, in scheme3, the carbonyl attached to R^(a) or R^(b) and the variable am convertedinto R⁴, and the other variable represents a suitable leaving group. Inscheme 4, R^(c) is converted into one of the moieties attached to thecarbonyl group within R⁴.

One route to compounds with the formula I using ketones is illustratedin Scheme 3. The intermediate 6 is obtained as illustrated in Scheme 1,followed by amide bond formation to afford the product of formula 7 asillustrated in Scheme 1. The intermediate 7 may be further elaborated byremoval of the Boc protecting group with, for example, trifluoroaceticacid, to give the unprotected secondary amine 8. This amine may befurther manipulated by the addition of a ketone R^(a)COR^(b) or aldehydeRCHO in a solvent such as 1,2-dichloroethane, in the presence of a mildacid, such as acetic acid, and a reducing agent, such as sodiumtriacetoxyborohydride, to form the alkylated amine 9.

The intermediate 8 may be otherwise manipulated by the methodillustrated in Scheme 4.

The amine 8 may be coupled to a carboxylic acid using1-ethyl-3-(3-dimethylainopropyl)carbodiimide (EDC),1-hydroxybenztriazole (HOBT), and a base, generally triethylamine ordiisopropylethylamine (DIPEA), in a solvent such asN,N-dimethylformamide (DMF), tetrahydrofuran (THF), or methylenechloride for 3 to 48 hours at ambient temperature to provideintermediate 10.

Intermediates such as 11 may also be synthesized by the general methodoutline in Scheme I from the corresponding ketones, which arecommercially available, known in the literature, or may be readilyprepared by those familiar with the art. In some cases intermediate 11may be deprotected using a mixture of aqueous hydrochloric acid and THFat 70° C. for 3 to 48 hours to afford the ketone intermediate 12, asillustrated in Scheme 5.

This intermediate may be further elaborated by the addition of a primaryamine, such as R¹¹NH₂, in dichloromethane in the presence of a mild acidsuch as acetic acid, and a reducing agent such as sodiumtriacetoxyborohydride, to afford the amine product 13. This intermediatemay be further elaborated by the addition of an aldeyde, also in thepresence of a reducing agent and a mild acid, to form the tertiary amineproduct 14. R^(a)CH— from the aldehyde form the variable R¹² or a groupthat is readily converted to R¹².

The intermediate 13 may alternatively be manipulated as illustrated inScheme 6, by the addition of a carboxylic acid R^(a)C(O)OH, using thepeptide coupling conditions utilized in Scheme 4, or by the addition ofan acid chloride, using the amide bond forming conditions utilized inScheme 1. This affords the amide product 15.

The following examples are illustrative of the present invention, andare not to be construed as limiting the scope of the appended claims.

EXAMPLE 1

tert-Butyl3-cyano-2-[(2-ethylbutanoyl)amino]-5,6-dihydrothieno[2,3-b]pyridine-7(4H)-carboxylate

Step A. tert-Butyl2-amino-3-cyano-5,6-dihydrothieno[2,3-b]pyridine-7(4H)-carboxylate

The tide compound was prepared via the sequence outlined in Scheme 1.Thus, to 0.200 g (1.00 mmol) of tert-butyl 3-oxopiperidine-1-carboxylatein 20 mL of EtOH was added 0.066 g (1.00 mmol) of malononitrile,followed by 0.088 mL (1.00 mmol) of morpholine, then 0.032 g (1.00 mmol)of elemental sulfur. The mixture was stirred at ambient temp for 16 h,then diluted with an equal volume of saturated aqueous NaHCO₃. Themixture was extracted twice with dichloromethane, and the combinedorganic layers were dried (Na₂SO₄) and concentrated in vacuo.Purification by flash chromatography (20% EtOAc in hexane) afforded thetide compound as a white solid.

¹H NMR (500 MHz, CDCl₃) 4.62 (s, 2H), 3.73 (t, J=5.5 Hz, 2H), 2.56 (t,J=6.5 Hz, 2H), 1.98 (m, 2H), 1.52 (8, 9H); mass spectrum (ES) m/e=280.2(M+H).

Step B. tert-Butyl3-cyano-2[(2-ethylbutanoyl)amino]-5,6-dihydrothienol[2.3-b]pyridine-7(4H)-carboxylate

To a solution of the material isolated in Step A in 1 mL ofdichloromethane was added 0.200 mL (1.15 mmol) ofdi-iso-propylethylamine, followed by 0.100 mL (0.728 mmol) of2-ethylbutanoyl chloride. After 16 hrs at ambient temperature themixture was diluted with 30 mL of dichloromethane, then washed twicewith an equal volume of saturated aqueous NaHCO₃. The organic layer wasdried (Na₂SO₄) and concentrated in vacuo, affording the title compound.

¹H NM (500 MHz, CDCl₃) 8.88 (s, 1H), 3.75 (s, 2H), 2.62 (t, J=6.0 Hz,2H), 2.30 (s, 1H), 1.99 (quint., J=5.5 Hz, 2H, 1.61 (m, 2H), 0.92 (t,J=7.0 Hz, 6H; mass spectrum (ES) m/e=378.2 (M+H).

EXAMPLE 2

N-(3-Cyano-7-isobutyl-4,5,6,7-tetrahydrothieno[2,3-b]pyridin-2-yl)-2-ethylbutanamide

Step A.N-(3-Cyano-4,5,6,7-tetrahydrothieno[2-3-b]pyridin-2-yl)-2-ethylbutanamide

To a solution of the tide compound from Example 1 in 20 mL ofdichloromethane was added 20 mL of trifluoroacetic acid. After 1 h atambient temperature, the mixture was concentrated in vacuo, and purifiedby flash chromatogaphy (40% EtOAc in hexane), affording the titlecompound.

¹H NMR (500 MHz, CDCl₃, TFA salt) 11.11 (s, 1H), 6.90 (s, 1H), 3.12 (s,1H), 2.46 (m, 2H), 1.78 (t, J=5.5 Hz, 2H), 1.50 (m, 2H), 1.42 (m, 2H),0.79 (t, J=7.5 Hz, 6H); mass spectrum (ES) m/e=278.2 (M+H).

Step B.N-(3-Cyano-7-isobutyl-4.5.6.7-tetrahydrothieno[2,3-b]pyridin-2-yl)-2-ethylbutanamide

To the material isolated in Step A in 2 mL of dichloromethane was added0.041 mL (0.90 mmol) of iso-butyraldehyde, followed by 0.026 mL (0.45mmol) of acetic acid and 0.10 g (0.45 mmol) of sodiumtracetoxyborohydride. After 16 h at ambient temperature the reaction wasdiluted with 30 mL of dichloromethane and washed twice with equalvolumes of saturated aqueous NaHCO₃. The organic layer was dried(Na₂SO₄) and concentrated in vacuo. Purification by preparative reversedphase BPLC afforded the tide compound.

¹H NMR (TFA salt) (500 MHz, CDCl₃) 8.90 (s, 1H), 3.13 (t, J=5.5 Hz, 1H),2.85 (d, J=7.5 Hz, 2H), 2.57 (t, J=6.5 Hz, 2H), 2.28 (m, 1H), 1.98 (m,2H), 1.72 (m, 2H), 1.59 (m, 21), 0.94 (t, J=7.5 Hz, 6H), 0.93 (t, J=6.5Hz, 6H); mass spectrum (ES) m/e=334 (M+1).

EXAMPLE 3

N-(3-Cyano-7-isopropyl-4,5,6,7-tetrahydrothieno[2,3-b]pyridin-2-yl)-2-ethylbutanamide

The title compound was prepared in an identical manner to Example 2.

¹H NMR (TFA salt) (500 Mz, CDCl₃) 8.56 (s, 1H): 3.64 (quint, J=7.0 Hz,1H), 3.07 (t, J=5.0 Hz, 2H), 2.57 (t, J=6.0 Hz, 2H), 2.46 (m, 1H), 1.97(quint, J=5.5 Hz, 2H), 1.74 (m, 2H), 1.60 (m, 2H), 1.19 (d, J=7.0 Hz,6H), 0.95 (t, J=7.5 Hz, 6H).

EXAMPLE 4

N-{6-[(4′-Chloro-1,1′-biphenyl-4-yl)methyl]-3-cyano-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl}-2-ethylbutanamide

Step A. tert-Butyl2-amino-3-cyano-4,7-dihythieno[2,3-c]pyridine-6-(5H)-carboxylate

The title compound was prepared via the sequence outlined in Scheme 1.Thus, to 10.0 g (50.2 mmol) of tert-butyl 4-oxopiperidine-1-carboxylatein 150 mL of EtOH was added 3.3 g (50.2 mmol) of malononitrile, followedby 6.6 mL (75 mmol) of morpholine, then 1.61 g (50.2 mmol) of elementalsulfur. The mixture was stirred at ambient temperature for 16 h, thenthe reaction mixture was filtered through a pad of silica, washing with50% EtOAc in hexane. The filtrate was concentrated in vacua, affordingthe title compound as a white solid.

¹H NMR (500 Mz, CDCl₃) 4.81 (s, 2H, 4.38 (s, 2H), 3.68 (t, J=5.0 Hz,2H), 2.61 (s, 2H), 1.50 (s, 9H); mass spectrum (ES) m/e=180.2 (M+H-Boc).

Step B. tert-Butyl3-cyano-2-[(2-ethylbutanoyl)amino]-4,7-dihydrothieno[2,3-c]pyridine-6(5H)-carboxylate

To a solution of 12.0 g (43.0 mmol) the intermediate prepared in Step Ain 250 mL of dichloromethane was added 15.0 mL (86 mmol) ofdi-iso-propylamine, followed by 9.0 mL (64 mmol) of 2-ethylbutanoylchloride. After 16 h at ambient temperature the reaction was dilutedwith an equal volume of saturated aqueous NaHCO₃, then extracted twicewith dichloromethane. The combined organic layers were dried (Na₂SO₄)and concentrated in vacuo. Purification by flash chromatography (15%EtOAc in hexane) afforded the title compound as a white solid.

¹H NMR (500 MHz, CDCl₃) 9.23 (s, 1H), 4.50 (s, 2H), 3.71 (s, 2H), 2.69(s, 2H), 2.35 (m, 1H), 1.76 (m, 2H), 1.63 (m, 2H), 1.50 (s, 9H), 0.95(t, J=7.5 Hz, 6H); mass spectrum (BS) m/e=378.1 (M+H).

Step C.N-(3-Cyano-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-2-ethylbutanamide

To a solution of the intermediate isolated in Step B in 100 mL ofdichloromethane was added 25 mL of trifluoroacetic acid. After 2 h atambient temperature, the reaction was diluted with an equal volume ofsaturated aqueous NAHCO₃. The mixture was extracted twice withdichloromethane, and the combined organic layers were dried (Na₂SO₄) andconcentrated in vacuo, affording the title compound.

¹H NMR (500 MHz, CDCl₃) 9.34 (s, 1H), 3.96 (s, 2H), 3.47 (s, 1H), 3.20(t, J=5.5 Hz, 2H, 2.67 (t, J=6.0 Hz, 2H, 2.33 (m, 1H), 1.73 (m, 2H),1.62 (m, 21p, 0.94 (s, 9H); mass spectrum (ES) m/e=278.2 (M+H).

Step D.N-{6-[(4′-Chloro-1,1′-biphenyl-4-yl)methyl]-3-cyano-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl}-2-ethylbutanamide

To a solution of the intermediate obtained in Step C in 2 mL of1,2-dichloroethane was added 98.0 mg (0.45 mmol) of4-(4-chlorophenyl)benzaldehyde, followed by 0.026 mL (0.45 mmol) ofacetic acid, and 100 mg (0.45 mmol) of sodium triacetoxyborohydride.After 16 h at ambient temperature the reaction was diluted with an equalvolume of saturated aqueous NaHCO₃, and the mixture was extracted twicewith dichloromethane. The combined organic layers were dried (Na₂SO₄)and concentrated in vacuo. Purification by reversed phase preparativeHPLC afforded the title compound as a white solid.

¹H NMR (TFA salt) (500 MHz, CDCl₃) 8.97 (s, 1H), 7.55 (m, 4H), 7.44 (m,4H), 3.78 (s, 2H), 3.61 (s, 2H), 2.89 (t, J=5.5 Hz, 2H), 2.75 (t, J=5.5Hz, 2H), 2.32 (m, 1H), 1.74 (m, 2H), 1.63 (m, 2H), 0.96 (t, J=7.0 Hz,6H); mass spectrum (ES) m/e=478.2 (M+H).

EXAMPLE 5

N-[3-Cyano-6-(4-phenoxybenzyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2-yl]-2-ethylbutanamide

The title compound was prepared in an identical manner to Example 4.

¹H NMR CIA salt) (500 MHz, CDCl₃) 9.26 (8, 1H), 7.35 (m, 4H), 7.13 (t,J=7.5 Hz, 1H), 7.04 (d, J=8.5 Hz, 2H), 7.01 (d, J=8.5 Hz, 2H), 3.71 (s,2H), 3.58 (s, 2H), 2.86 (t, J=5.5 Hz, 2H, 2.74 (t, J=5.0 Hz, 2H), 2.37(m, 1H), 1.75 (m, 2H), 1.62 (m, 2H), 0.96 (t, J=7.5 Hz, 6H); massspectrum (ES) m/e=460.1 (M+H).

EXAMPLE 6

N-{6-[4-(4-Chlorophenoxy)benzyl]-3-cyano-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2-yl}-2-ethylbutanamide

The title compound was prepared in an identical manner to Example 4.

¹H NMR (TFA salt) (500 Mhz, CDCl₃) 9.13 (s, 1H), 7.34 (t, J=8.0 Hz, 2H),7.30 (d, J=8.5 Hz, 2H), 6.97 (d, J=8.5H, 2H), 6.96 (d, J=9.5 Hz, 2H),3.70 (s, 2H), 3.57 (s, 2H), 2.85 (t, J=6.0 Hz, 2H), 2.72 (t, J=5.5 Hz,2H), 2.33 (m, 1H), 1.75 (m, 2H), 1.61 (m, 2H), 0.94 (t, J=7.5 Hz, 6H);mass spectrum (ES) m/e=494.2 (M+H).

EXAMPLE 7

N-[3-Cyano-6-(3-phenoxybenzyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2-yl]-2-ethylbutanamide

The title compound was prepared in an identical manner to Example 4.

¹H NMR (TFA salt) (500 MHz, CDCl₃) 9.21 (s, 1H), 7.34 (m, 3H), 7.13 (d,J=7.5 Hz, 2H), 7.07 (s, 1H), 7.03 (d, J=8.0 Hz, 2H), 6.95 (d, J=8.0 Hz,1H), 3.73 (s, 2H), 3.60 (s, 2H), 2.86 (t, J=5.0 Hz, 2H), 2.72 (t, J=4.5Hz, 2H), 2.35 (m, 1H), 1.75 (m, 2H), 1.62 (m, 2H), 0.96 (t, J=7.5 Hz,6H); mass spectrum (ES) m/e=460.1 (M+H).

EXAMPLE 8

N-(3-Cyano-6-{[1-(2,4-dichlorophenyl)cylopropyl]carbonyl}-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-2-ethylbutanamide

To a solution of the intermediate prepared in Step C of Example 4 in 1mL of DMP was added 4.2 mg (0.14 mmol) of1-(2,4-dichlorophenyl)cyclopropanecarboxylic acid, followed by 0.047 mL(0.027 mmol) of di-isopropylethylamine and 68.0 mg (0.18 mmol) ofO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramehyluroniumhexafluorophosphate (HATU). After 16 h at ambient temperature themixture was diluted with 50 mL of EtOAc. The organic layer was washedwith saturated aqueous NaICO₃, and brine, dried (MgSO₄) and concentratedin vacuo. Purification by flash chromatography (30% EtOAc in hexane)afforded the tide compound as a white solid.

¹H NMR (500 MHz, CDCl₃) 8.69 (s, 1H), 7.38 (s, 1H), 7.29 (s, 2H), 4.46(s, 2H), 3.71 (m, 2H), 2.26 (m, 1H), 1.74 (m, 2H), 1.68 (q, J=2.5 Hz,2H), 1.62 (m, 2H), 1.16 (q, J=2.5 Hz, 2H), 0.93 (t, J=7.5 Hz, 6H); massspectrum (ES) m/e=490.1 (M+H).

EXAMPLE 9

N-{3-Cyano-6[(2,4-dichlorobezyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide

Step A. The title compound was prepared via the sequence outlined inScheme 1. Thus, to 1,4-dioxaspiro(4.5]decan-8-one in 200 mL of EtOH wasadded 10.1 g (160 mmol) of malononitrile, followed by 22.0 mL (240 mmol)of morpholine, then 5.13 g (160 mmol) of elemental sulfur. The mixtureheated to 100° C. for 16 h, then diluted with an equal volume ofsaturated aqueous NaHCO₃. The mixture was extracted twice withdichloromethane, and the combined organic layers were dried (Na₂SO₄) andconcentrated in vacuo, affording a yellow solid.

Mass spectrum (ES) m/e=237.1 (M+H).

The crude material was carried directly on to Step B.

Step B. The crude material from Step A was dissolved in 200 mL ofdichloromethane. To this solution was added 46.0 mL (263 mmol) ofdi-iso-propylethylamine, followed by 8.91 mL (66.0 mmol) of2-ethylbutanoyl chloride. After 16 h at ambient temperature, thereaction was quenched with an equal volume of saturated aqueous NaHCO₃.The mixture was extracted twice with dichloromethane, and the combinedorganic layers were dried (Na₂SO₄) and concentrated in vacuo, affordinga yellow solid.

Mass spectrum (ES) m/e=335.2 (M+H).

The crude material was carried directly on to Step C.

Step C.N-(3-Cyano-6-oxo-4,5,6,7-tetrahydro-1-benzothien-2-yl)-2-ethylbutanamide

To the material isolated in Step B in 100 mL of THF was added 52 mL (52mmol) of 1.0 N aqueous HCl. The mixture was heated to 70° C. for 48 h,then cooled to ambient temperature and added to 52 mL (52 mmol) of 1.0 NNaOH. The resulting mixture was extracted twice with dichloromethane,and the combined organic layers were dried (Na₂SO₄) and concentrated invacuo. Purification by flash chromatography (20% EtOAc in hexane)afforded 0.650 g of the intermediate as a white solid.

¹H NMR (500 MHz, CDCl₃) 9.17 (s, 1H), 3.52 (s, 2H), 3.03 (t, J=6.5 Hz,2H), 2.72 (t, J=8.0 Hz, 2H), 2.36 (m, 1H), 1.77 (m, 2H), 1.65 (m, 2H),0.95 (t, J=7.5 Hz, 6H); mass spectrum (ES) m/e=291.2 (M+H).

Step D.N-{3-Cyano-6-[(2,4-dichlorobenzyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide

To a solution of the intermediate isolated in Step C in 100 mL ofdichloromethane was added 1.41 mL (12.0 mL) of1-(2,4-dichlorophenyl)methanamine, followed by 2.54 g (12.0 mmol) ofsodium triacetoxyborohydride, and 0.960 mL (16.0 mmol) of acetic acid.After 16 h at ambient temperature, the reaction was quenched with anequal volume of saturated aqueous NaHCO₃. The mixture was extractedtwice with dichloromethane, and the combined organic layers were dried(Na₂SO₄), and concentrated in vacuo, affording a yellow solid.Purification by flash chromatography (15% EtOAc in hexane) afforded thetitle compound as a white solid.

¹H NMR (500 MHz, CDCl₃) 8.54 (s, 1H), 7.39 (m, 2H, 7.25 (m, 1H), 3.95(s, 2H), 3.04 (m, 1H), 2.95 (dd, J=4.0 Hz, J=16.0 Hz, 1H), 2.78 (dt,J=5.5 Hz, J=16.5 Hz, 1H), 2.62 (m, 1H), 2.53 (dd, J=8.0 Hz, J=16.0 Hz,1H), 2.25 (m, 1H), 2.09 (m, 1H), 1.76 (m, 2H), 1.64 (m, 2H), 1.56 (8,1H), 0.95 (t, J=7.0 Hz); mas spectrum (ES) m/e=450.1 (M+H).

EXAMPLE 10

N-{3-Cyano-6-[cyclopropylmethyl)(2,4-dichlorobenzyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide

To a solution of the title compound from Example 9 in 5 mL ofdichloromethane was added 0.165 mL (2.20 mmol) ofcyclopropanecarbaldehyde, followed by 0.187 g (0.88 mmol) of sodiumtriacetoxyborohydride. After 72 h at ambient temperature the mixture wasdiluted with an equal volume of saturated aqueous NaHCO₃, and extractedtwice with dichloromethane. The combined organic layers were dried(Na₂SO₄) and concentrated in vacuo. Purification by flashchromatography, followed by purification by reversed phase preparativeHPLC, afforded the title compound as a white solid.

¹H NMR (TFA salt) (500 MHz, CDCl₃) 8.28 (s, 1H), 7.74 (s, 1H), 7.38 (s,1H), 7.29 (s, 1H), 3.99 (s, 2H), 2.91 (m, 1H), 2.85 (dd, J=7.0 Hz,J=17.0 Hz, 1H), 2.75 (m, 1H), 2.60 (m, 2H), 2.23 (m, 1H), 1.70 (m, 2H),1.63 (m, 21), 0.95 (t, J=8.0 Hz, 6); mass spectrum (ES) m/e=504.1 (M+H).

Using the title compound prepared in Example 9, and following theprocedure outlined in Example 10, the compounds listed in Table 3 wereprepared.

TABLE 3

Example R¹² Mass spectrum (ES) m/e = 11

492.1 (M + H) 12

520.2 (M + H) 13

534.2 (M + H) 14

506.2 (M + H) 15

534.2 (M + H) 16

450.1 (M + H) 17

568.2 (M + H) 18

580.2 (M + H) 19

346.2 (M + H) 20

558.1 (M + H) 21

534.2 (M + H) 22

544.2 (M + H) 23

633.2 (M + H)

Using the intermediate prepared in Example 9 Step C, and the procedureutilized in Example 9 Step D, the compounds listed in Table 4 wereprepared.

TABLE 4

Mass spectrum Example R⁵ (ES) m/e = 24

518.3 (M + H) 25

465.3 (M + H) 26

465.3 (M + H) 27

422.3 (M + H) 28

422.3 (M + H) 29

416.3 (M + H) 30

476.2 (M + H) 31

422.3 (M + H) 32

437.2 (M + H)

EXAMPLE 33

N-{3-Cyano-6-[(3,4-dichlorobenzyl)(methyl)amino]-4,5,6,7-tetrahydro-1-benzothien-2-yl}-2-ethylbutanamide

To a solution of the title compound from Example 24 in 2 mL ofdichloromethane was added 0.060 mL (0.65 mmol) of 37% aqueousformaldehyde, followed by 0.055 g (0.26 mmol) of sodiumtriacetoxyborohydride. After 16 h at ambient temperature, the reactionwas diluted with 30 mL of dichloromethane and washed twice with an equalvolume of saturated aqueous NaHCO₃. The organic layer was dried (Na₂SO₄)and concentrated in vacuo. Purification by preparative reversed phaseHPLC afforded the title compound as a white solid (TFA salt).

¹H NMR (500 MHz, CDCl₃) 8.64 (s, 1H), 7.64 (m, 1H), 7.41 (m, 1H), 7.14(d, J=8.0 Hz, 1H), 6.07 (s, 1H), 4.42 (d, J=5.5 Hz, 2H), 4.26 (m, 1H),3.66 (m, 1H), 2.98 (m, 2H), 2.79 (s, 3H), 2.68 (m, 2H), 2.33 (m, 1H),2.00 (m, 1H), 1.72 (m, 2H), 1.62 (m, 2H), 0.95 (t, J=8.0 Hz, 6H; massspectrum (ES) m/e=464.2 (M+H).

Utilizing the procedure outlined in Example 33, and the title compoundfrom the indicated Example as starting material, the compounds listed inTable 5 were prepared.

TABLE 5

S. Mat. Mass Spectrum Example (Example) R¹¹ (ES) m/e = 34 25

479.3 (M + H) 35 26

479.3 (M + H) 36 27

436.3 (M + H) 37 28

436.3 (M + H) 38 29

430.3 (M + H) 39 30

490.2 (M + H)

EXAMPLE 40

N-{3-Cyano-2-[(2-ethylbutanoyl)amino]-4,5,6,7-tetrahydro-1-benzothien-6-yl}-N-[1-hydroxymethyl)-2,2-dimethylpropyl]cyclohexanecarboxamideStep A.N-(3-Cyano-6-{[1-(hydroxymethyl)-2,2-dimethylpropyl]amino}-4,5,6,7-tetrahydro-1-benzothien-2-yl)-2-ethylbutanamide

To a solution of the intermediate isolated in Example 9, Step C, in 40mL of dichloromethane was added 0.452 g (3.90 mmol) of2-amino-3,3-dimethylbutan-1-ol, followed by 1.24 g (5.9 mmol) of sodiumtriacetoxyborohydride and 0.468 mL (7.80 mmol) of acetic acid. After 16h at ambient temperature the reaction was diluted with an equal volumeof saturated aqueous NaHCO₃. The organic layer was dried (Na₂SO₄) andconcentrated in vacuo. Purification by preparative reversed phase HPLCafforded the title compound (TFA salt).

Mass spectrum (ES) m/e=392.3 (M+H).

Step B.N-{3-Cyano-2-[(2-ethylbutanoyl)amino]-4,5,6,7-tetrahydro-1-benzothien-6-yl}-N-[1-(hydroxymethyl)-2,2-dimethylpropyl]cyclohexanecarboxamide

To a solution of the intermediate isolated in Step A in 5.0 mL ofdichloromethane was added 0.175 mL (1.00 mmol) of di-isopropylethylamineand 0.041 mL (0.280 mmol) of cyclohexanecarbonyl chloride. Afterstirring 16 h at ambient temperate, the reaction mixture wasconcentrated in vacuo, and purified by preparative reversed phase HPLC,affording the title compound as a white solid.

¹H NMR (500 MHz, CDCl₃) 8.51 (s, 1H), 4.52 (dt, J=3.5 Hz, J=12.5 Hz,1H), 4.42 (dd, J=7.5 Hz, J=13.0 Hz, 1H), 3.75 (m, 1H), 3.08 (m, 2H),2.88 (m, 1H), 2.67 (m, 1H), 2.51 (m, 1H), 2.27 (m, 3H), 2.18 (m, 1H),1.86 (d, J=11.0 Hz, 1H), 1.76 (m, 3H), 1.62 (m, 2H), 1.40 (m, 1H), 1.27(m, 1H), 1.20 (s, 9H), 0.95 (t, J=7.0 Hz); mass spectrum (ES) m/e=502.4(M+H).

Utilizing the procedure outlined in Example 40 Step B, and the startingmaterial synthesized in Example 40, Step A, the compounds listed inTable 6 were synthesized.

TABLE 6

Example R¹² Mass spectrum (ES) m/e = 41

460.3 (M + H) 42

490.4 (M + H) 43

488.4 (M + H) 44

496.4 (M + H)

EXAMPLE 45

N-{3-Cyano-2-[(2-ethylbutanoyl)amino]-4,5,6,7-tetrahydro-1-benzothien-6-yl}-N-(2,4-dichlorobenzyl)-3,3-dimethylbutanamide

To a solution of the title compound from Example 9 in 50 mL ofdichloromethane was added 0.307 mL (1.80 mmol) ofdi-iso-propylethylamine, a catalytic amount (ca. 2 mg) of HOBT, and0.065 mL (0.48 mmol) of 3,3-ethylbutanoyl chloride. After 16 h atambient temperature the read on was diluted with an equal volume ofsaturated aqueous NaHCO₃. The organic layer was dried (NSO₄) andconcentrated in vacuo. Purification by preparative reversed phase HPLCafforded the title compound as a white solid.

¹HNMR (300 MHz, CDCl₃) 8.52 (s, 1H), 7.46 (s, 1H), 7.34 (m, 1H), 7.20(m, 1H), 4.97 (m, 1 μl), 4.72 (d, J=17 Hz, 1H), 4.59 (d, J=16 Hz, 1H),2.73 (m, 4H), 2.25 (m, 1H), 2.12 (s, 2H, 1.86 (m, 2H), 1.78 (m, 2H),1.60 (m, 2H), 1.07 (s, 9H), 0.94 (t, J=7.5 Hz, 6H); mass spectrum (ES)m/e=548.1 (M+H).

Biological Assays

The ability of the compounds of the present invention to inhibit thebinding of glucagon can be demonstrated using the following in vitroassays.

Glucapon Receptor Binding Assay

A stable CHO (Chinese hamster Ovary) cell line expressing cloned humanglucagon receptor was maintained as described (Chicchi et al. J BiolChem 272, 7765-9(1997); Cascieri et al. J Biol Chem 274, 8694-7(1999)).To determine antagonistic binding affinity of compounds 0.002 mg of cellmembranes from these cells were incubated with ¹²⁵I-Glucagon (NewEngland Nuclear, MA) in a buffer containing 50 mM Tris-HCl (pH 7.5), 5mM MgCl₂, 2 mM EDTA, 12% Glycerol, and 0.200 mg WGA coated PVT SPA beads(Amersham), +/− compounds or 0.001 mM unlabeled glucagon. After 412hours incubation at room temperature, the radioactivity bound to thecell membranes was determined in a radioactive emission detectioncounter (Mcrobeta-Wallace). Data was analyzed using the software programPrisms from GraphPad. The IC₅₀ were calculated using non-linearregression analysis assuming single site competition.

High Throughput Screening (HTS) Protocol for Glucagon Receptor BindingAssay

Another form of the binding assay was developed suitable forhigh-throughput screening for modulators of receptor activity. Fullyautomated or semi-automated protocols and robotic and workstationinstruments were utilized for the HTS assay as would be recognized bythose practiced in the art. In a typical configuration of the assay,0.002 mg of cell membrane (as described above) were preincubated with0.200 mg of WGA-coated PVT beads in buffer containing 100 mM Tris-HCl pH7.5, 10 mM MgCl₂, 4 mM EDTA, 24% Glycerol, and 0.2% BSA. Themembrane/bead mixture was then dispensed (0.050 mL) into each well of96-well plates (Wallac Isoplates, white clear bottom) containing 0.100mL of test compounds or control solutions. A second addition (0.050 mL)was then dispensed into the wells of the plate containing ¹²⁵I-Glucagon(approximately 25,000 CPM). The solutions were dispensed using aMultidrop Stacker 20 (Titertek) liquid dispenser. An adhesive plate seal(Packard) was applied and the plates were shaken for 5 minutes. Theplates were further incubated at ambient temperature for several hoursfor establishment of equilibrium (typically 5 hours) and the signal wasstable for up to three days. The plates were read in a scintillationcounter (Wallac Microbeta) for 1 min/well. Activity of test compoundswas calculated by comparing to the total scintillation signal (CPM) ofcontrol samples with no compound and with 0.001 mM unlabeled-glucagon.

Inhibition of Glucagon-stimulated Intracellular cAMP Formation

Exponentially growing CHO cells expressing human glucagon receptor wereharvested with the aid of enzyme-free dissociation media (SpecialtyMedia), pelleted at low speed, and re-suspended in cell suspensionbuffer [75 mM Tris-HCl pH 7.5, 250 mM Sucrose, 25 mM MgCl₂, 1.5 mM EDTA,0.1 mM Ro-20-1724 (Biomol, Inc.), 0.2% bovine serum albumin and onetablet of complete™ (Boehringer), which contains a cocktail of proteaseinhibitors, for each 50 ml of buffer]. An adenylate cyclase assay wassetup using an Adenylate Cyclase Assay kit (SMP-004B) from New EnglandNuclear (NEN) as per manufacturer instructions. Briefly, compounds werediluted from stocks in a cell stimulation buffer supplied with the kit.Cells prepared as above were preincubated in flash plates coated withanti-cAMP antibodies (NEN) in presence of compounds or DMSO controls for40 minutes, and then stimulated with glucagon (250 pM) for an additional40 minutes. The cell stimulation was stopped by addition of equal amountof a detection buffer containing lysis buffer as well as ¹²⁵I-labeledcAMP tracer (NEN). After 3-6 h of incubation at room temperature thebound radioactivity was determined in a liquid scintillation counterTopCount-Packard Instruments). Activity of test compounds was calculatedby comparing to the total scintillation signal (CPM) of control sampleswith no compound and with 0.001 mM unlabeled-glucagon.

Certain embodiments of the invention has been described in detail;however, numerous other embodiments are contemplated as falling withinthe invention. Thus, the claims are not limited to the specificembodiments described herein. All patents, patent applications andpublications that are cited herein are hereby incorporated by referencein their entirety.

1. A compound represented by formula I:

or a pharmaceutically acceptable salt or solvate thereof wherein: X isNR⁴; R¹ is selected from the group consisting of: H, C₁₋₁₀alkyl,C₃₋₇cycloalkyl and Aryl, said alkyl, cycloalkyl and Aryl beingoptionally substituted with 1-4 substituents independently selected fromR¹³; R² is selected from the group consisting of: R¹ as defined above,—C(O)₂R⁷ and —CONR⁷R⁸; m is 0; n is 3; R³ is selected from the groupconsisting of: C₁₋₁₀alkyl, C₃₋₇cycloalkyl and Aryl, said alkyl,cycloalkyl and Aryl being optionally substituted with 1-4 substituentsselected from R¹³, such that when R³ represents C₁₋₁₀ alkyl substitutedwith one R¹³ group, and R¹³ represents halo, R¹, R², R⁵ and R⁶ do notrepresent C₁₋₃alkyl; R⁴ is selected from the group consisting of: C₃₋₁₀alkyl, C₃₋₇ cycloalkyl, Aryl, HAR, Hetcy, C(O)C₅₋₁₀ alkyl, C(O)C₃₋₇cycloalkyl, C(O)-Aryl, C(O)-HAR, C(O)-Hetcy, CONR⁹R¹⁰, CO₂R⁹ and SO₂R⁹,the alkyl, cycloalkyl, Aryl, HAR and Hetcy groups and portions beingoptionally substituted with 1-4 substituents selected from R¹³; one ofR⁵ and R⁶ is selected from the group consisting of NR¹¹R¹², NR¹¹COR¹²,NR¹¹CO₂R¹² and NR¹¹S(O)₂R¹², and the other represents R¹, HAR, Hetcy orOR¹¹, said HAR and Hetcy being optionally substituted with 1-4substituents selected from R¹³, R⁷, R¹⁰ and R¹¹ are selected from thegroup consisting of: R¹ as defined above, HAR and Hetcy, said HAR andHetcy being optionally substituted with 1-4 substituents selected fromR¹³; R⁸, R⁹ and R¹² are selected from the group consisting of:C₁₋₁₀alkyl, C₃₋₇cycloalkyl, Aryl, HAR and Hetcy, said alkyl, cycloalkyl,Aryl, HAR and Hetcy being optionally substituted with 1-4 substituentsselected from R¹³; or alternatively, R⁷, R⁸, R⁹ and R¹⁰ are as definedabove, and R¹¹ and R¹² are taken together with the atoms to which theyare attached along with any intervening atoms and represent a 5-8membered ring optionally containing 1-2 heteroatoms selected from O, Sand N, and optionally substituted with 1-4 substituents selected fromR¹³; each R¹³ is selected from the group consisting of: halo, NR¹⁴R¹⁵,C₁₋₄alkyl, C₃₋₇ cycloalkyl, Aryl, HAR, Hetcy, CF₃, OCF₃, OR¹⁵, NO₂,S(O)_(x)R¹⁴, SR¹⁴, S(O)_(x)NR¹⁴R¹⁵, O(CR¹⁶R¹⁷)_(y)NR¹⁴R¹⁵, C(O)R¹⁴,CO₂R¹⁵, CO₂(CR¹⁶R¹⁷)yCONR¹⁴R¹⁵, OC(O)R¹⁴, CN, C(O)NR¹⁴R¹⁵, NR¹⁵C(O)R¹⁴,NR¹⁵C(O)OR¹⁴, NR¹⁵C(O)NR¹⁶R¹⁴ and CR¹⁵(N—OR¹⁴), wherein x is 1 or 2, andy is an integer from 1-4, said alkyl, cycloalkyl, Aryl, HAR and Hetcybeing optionally substituted with 1-4 substituents selected from R¹⁸;R¹⁴, R¹⁵, R¹⁶ and R¹⁷ are independently selected from the groupconsisting of: H, C₁₋₁₀alkyl, C₃₋₇cycloalkyl, Aryl and Ar—C₁₋₁₀alkyl;and each R¹⁸ is independently selected from the group consisting of:halogen, CN, C₁₋₄alkyl, OH, CF₃, Aryl, Aryloxy, CO₂H and CO₂C₁₋₄ alkyl,said Aryl and the Aryl portion of Aryloxy being optionally substitutedwith up to 4 halo groups, and up to 2 C₁₋₄ alkyl, OH, CF₃ or CN groups.2. A compound in accordance with claim 1 wherein R¹ is selected from thegroup consisting of: H, C₁₋₁₀alkyl, C₃₋₆ cycloalkyl and phenyl, saidalkyl and phenyl being optionally substituted with 1-3 substituentsselected from R¹³.
 3. A compound in accordance with claim 1 wherein R²is H.
 4. A compound in accordance with claim 1 wherein R³ is C₃₋₁₀ alkyloptionally substituted with 1-4 substituents selected from R¹³, suchthat when R³ is substituted with one R¹³ group, and R¹³ represents halo,R¹, R², R¹ and R⁶ do not represent C₁₋₃alkyl.
 5. A compound inaccordance with claim 4 wherein R³ represents C₃₋₅ alkyl, optionallysubstituted with 1-4 R¹³ groups.
 6. A compound in accordance with claim1 wherein R⁴ is selected from the group consisting of: C₅s₁₀ alkyl, C₃₋₆cycloalkyl, phenyl, HAR, Hetcy, C(O)C₅₋₁₀alkyl, C(O)C₃₋₆ cycloalkyl andCO₂ ⁹, the alkyl, cycloalkyl and, Aryl groups and portions, phenyl, HARand Hetcy being optionally substituted with 1-4 substituents selectedfrom R¹³, and R⁹ representing C₁₋₁₀alkyl, C₃₋₇cycloalkyl, Aryl, HAR orHetcy, said alkyl, cycloalkyl, Aryl groups and portions, HAR and Hetcybeing optionally substituted with 1-4 R¹³ groups.
 7. A compound inaccordance with claim 1 wherein R¹³ is selected from the groupconsisting of: halo, C₁₋₄alkyl, C₃₋₇cycloalkyl, Aryl, HAR, Hetcy, andOR¹⁵ wherein R¹⁵ is H, said alkyl, cycloalkyl, Aryl, HAR and Hetcy beingoptionally substituted with 1-4 substituents selected from R¹⁸ and R¹⁸is halo, C₁₋₄alkyl, Aryl or CO₂C₁₋₄ alkyl.
 8. A compound in accordancewith claim 1 selected from the group consisting of: tert-butyl3-cyano-2-[(2-ethylbutanoyl)amino]-5,6-dihydrothieno[2,3-b]pyridine-7(4H)carboxylate;N-(3-cyano-7-isobutyl-4,5,6,7-tetrahydrothieno[2,3-b]pyridin-2-yl)-2-ethylbutanamide;andN-(3-cyano-7-isopropyl-4,5,6,7-tetrahydrothieno[2,3-b]pyridin-2-yl)-2-ethylbutanamide.9. A pharmaceutical composition which is comprised of a compound inaccordance with claim 1 in combination with a pharmaceuticallyacceptable carrier.
 10. A method of treating type 2 diabetes mellitus ina mammalian patient in need of such treatment, comprising administeringto said patient a compound in accordance with claim 1 in an amount thatis effective to treat type 2 diabetes mellitus.